Non-coding RNAs and Mechanisms of Cytoplasmic Gene Regulation, Berlin Institute for Medical Systems Biology, Max Delbrück Center for Molecular Medicine, Berlin, Germany.
Non-coding RNAs and Mechanisms of Cytoplasmic Gene Regulation, Berlin Institute for Medical Systems Biology, Max Delbrück Center for Molecular Medicine, Berlin, Germany.
Methods. 2019 Jun 1;162-163:31-41. doi: 10.1016/j.ymeth.2019.02.002. Epub 2019 Feb 8.
The subcellular localization and translation of mRNAs are fundamental biological processes. In neurons, they underlie cell growth and synaptic plasticity, which serves as a foundation of learning and memory. Multiple approaches have been developed to separate neurons on subcellular compartments - cell bodies (soma) and cell extensions (axons and dendrites) - for further biochemical analyses. Here we describe neurite/soma separation approach in combination with RNA sequencing and proteomic analyses to identify localized and locally translated RNAs and proteins. This approach allows quantification of around 7000 of local proteins and the entire local transcriptome. It provides a powerful tool for investigation of the mechanisms underlying RNA localization and local translation in neurons.
mRNA 的亚细胞定位和翻译是基本的生物学过程。在神经元中，它们是细胞生长和突触可塑性的基础，而后者是学习和记忆的基础。已经开发出多种方法来分离神经元的亚细胞区室 - 细胞体（soma）和细胞延伸（轴突和树突） - 以进行进一步的生化分析。在这里，我们描述了与 RNA 测序和蛋白质组学分析相结合的轴突/细胞体分离方法，以鉴定局部和局部翻译的 RNA 和蛋白质。该方法可以定量大约 7000 种局部蛋白质和整个局部转录组。它为研究神经元中 RNA 定位和局部翻译的机制提供了有力的工具。
